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Detection of apoptosis-related factors in NHEK cell lysates following sorafenib and antifungal drug treatment. Detection of (A) cIAP-1 (70 kDa) and (C) cleaved caspase-3 (19 kDa) in the cell lysate was performed using western blotting. (B) Bands were semi-quantified using ImageJ, and data were expressed as the relative ratio of cIAP-1/β-actin. Statistical analysis was conducted using one-way ANOVA followed by Tukey’s test; ns indicates not significant. Sora or S: sorafenib, Keto: ketoconazole, Itra: itraconazole. All results are presented as the mean ± SD of triplicate experiments. (D) Various concentrations of sorafenib, with or without 50 μmM ZVAD or <t>ZDEVD,</t> were added to cultured NHEK cells. DMSO was used as the solvent for both sorafenib and the inhibitors. After overnight incubation, cell viability was assessed using the cell-counting kit-8 assay. Statistical analysis was performed using one-way ANOVA followed by Dunnett’s test; ** p < 0.01 and * p < 0.05 (vs DMSO (-)).
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Detection of apoptosis-related factors in NHEK cell lysates following sorafenib and antifungal drug treatment. Detection of (A) cIAP-1 (70 kDa) and (C) cleaved caspase-3 (19 kDa) in the cell lysate was performed using western blotting. (B) Bands were semi-quantified using ImageJ, and data were expressed as the relative ratio of cIAP-1/β-actin. Statistical analysis was conducted using one-way ANOVA followed by Tukey’s test; ns indicates not significant. Sora or S: sorafenib, Keto: ketoconazole, Itra: itraconazole. All results are presented as the mean ± SD of triplicate experiments. (D) Various concentrations of sorafenib, with or without 50 μmM ZVAD or <t>ZDEVD,</t> were added to cultured NHEK cells. DMSO was used as the solvent for both sorafenib and the inhibitors. After overnight incubation, cell viability was assessed using the cell-counting kit-8 assay. Statistical analysis was performed using one-way ANOVA followed by Dunnett’s test; ** p < 0.01 and * p < 0.05 (vs DMSO (-)).
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Detection of apoptosis-related factors in NHEK cell lysates following sorafenib and antifungal drug treatment. Detection of (A) cIAP-1 (70 kDa) and (C) cleaved caspase-3 (19 kDa) in the cell lysate was performed using western blotting. (B) Bands were semi-quantified using ImageJ, and data were expressed as the relative ratio of cIAP-1/β-actin. Statistical analysis was conducted using one-way ANOVA followed by Tukey’s test; ns indicates not significant. Sora or S: sorafenib, Keto: ketoconazole, Itra: itraconazole. All results are presented as the mean ± SD of triplicate experiments. (D) Various concentrations of sorafenib, with or without 50 μmM ZVAD or <t>ZDEVD,</t> were added to cultured NHEK cells. DMSO was used as the solvent for both sorafenib and the inhibitors. After overnight incubation, cell viability was assessed using the cell-counting kit-8 assay. Statistical analysis was performed using one-way ANOVA followed by Dunnett’s test; ** p < 0.01 and * p < 0.05 (vs DMSO (-)).
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Detection of apoptosis-related factors in NHEK cell lysates following sorafenib and antifungal drug treatment. Detection of (A) cIAP-1 (70 kDa) and (C) cleaved caspase-3 (19 kDa) in the cell lysate was performed using western blotting. (B) Bands were semi-quantified using ImageJ, and data were expressed as the relative ratio of cIAP-1/β-actin. Statistical analysis was conducted using one-way ANOVA followed by Tukey’s test; ns indicates not significant. Sora or S: sorafenib, Keto: ketoconazole, Itra: itraconazole. All results are presented as the mean ± SD of triplicate experiments. (D) Various concentrations of sorafenib, with or without 50 μmM ZVAD or <t>ZDEVD,</t> were added to cultured NHEK cells. DMSO was used as the solvent for both sorafenib and the inhibitors. After overnight incubation, cell viability was assessed using the cell-counting kit-8 assay. Statistical analysis was performed using one-way ANOVA followed by Dunnett’s test; ** p < 0.01 and * p < 0.05 (vs DMSO (-)).
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Detection of apoptosis-related factors in NHEK cell lysates following sorafenib and antifungal drug treatment. Detection of (A) cIAP-1 (70 kDa) and (C) cleaved caspase-3 (19 kDa) in the cell lysate was performed using western blotting. (B) Bands were semi-quantified using ImageJ, and data were expressed as the relative ratio of cIAP-1/β-actin. Statistical analysis was conducted using one-way ANOVA followed by Tukey’s test; ns indicates not significant. Sora or S: sorafenib, Keto: ketoconazole, Itra: itraconazole. All results are presented as the mean ± SD of triplicate experiments. (D) Various concentrations of sorafenib, with or without 50 μmM ZVAD or <t>ZDEVD,</t> were added to cultured NHEK cells. DMSO was used as the solvent for both sorafenib and the inhibitors. After overnight incubation, cell viability was assessed using the cell-counting kit-8 assay. Statistical analysis was performed using one-way ANOVA followed by Dunnett’s test; ** p < 0.01 and * p < 0.05 (vs DMSO (-)).
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Detection of apoptosis-related factors in NHEK cell lysates following sorafenib and antifungal drug treatment. Detection of (A) cIAP-1 (70 kDa) and (C) cleaved caspase-3 (19 kDa) in the cell lysate was performed using western blotting. (B) Bands were semi-quantified using ImageJ, and data were expressed as the relative ratio of cIAP-1/β-actin. Statistical analysis was conducted using one-way ANOVA followed by Tukey’s test; ns indicates not significant. Sora or S: sorafenib, Keto: ketoconazole, Itra: itraconazole. All results are presented as the mean ± SD of triplicate experiments. (D) Various concentrations of sorafenib, with or without 50 μmM ZVAD or ZDEVD, were added to cultured NHEK cells. DMSO was used as the solvent for both sorafenib and the inhibitors. After overnight incubation, cell viability was assessed using the cell-counting kit-8 assay. Statistical analysis was performed using one-way ANOVA followed by Dunnett’s test; ** p < 0.01 and * p < 0.05 (vs DMSO (-)).

Journal: Acta Dermato-Venereologica

Article Title: Possible Clinical Effects of Ketoconazole on Sorafenib-induced Hand–Foot Skin Reaction and Cytoprotection Mechanisms of Antifungal Agents against Multikinase Inhibitor-induced Keratinocyte Toxicity

doi: 10.2340/actadv.v105.40697

Figure Lengend Snippet: Detection of apoptosis-related factors in NHEK cell lysates following sorafenib and antifungal drug treatment. Detection of (A) cIAP-1 (70 kDa) and (C) cleaved caspase-3 (19 kDa) in the cell lysate was performed using western blotting. (B) Bands were semi-quantified using ImageJ, and data were expressed as the relative ratio of cIAP-1/β-actin. Statistical analysis was conducted using one-way ANOVA followed by Tukey’s test; ns indicates not significant. Sora or S: sorafenib, Keto: ketoconazole, Itra: itraconazole. All results are presented as the mean ± SD of triplicate experiments. (D) Various concentrations of sorafenib, with or without 50 μmM ZVAD or ZDEVD, were added to cultured NHEK cells. DMSO was used as the solvent for both sorafenib and the inhibitors. After overnight incubation, cell viability was assessed using the cell-counting kit-8 assay. Statistical analysis was performed using one-way ANOVA followed by Dunnett’s test; ** p < 0.01 and * p < 0.05 (vs DMSO (-)).

Article Snippet: Z-DEVD-FMK (ZDEVD) and Z-VAD-FMK (ZVAD) were from Selleck Chemicals (Houston, TX, USA).

Techniques: Western Blot, Cell Culture, Solvent, Incubation, Cell Counting